Oxygen Gradient Ektacytometry Is Associated with Markers of Hemolysis and Inflammation in a Large Sickle Cell Disease Cohort within the GenoMed4ALL Project

Idrizovic, Amira

Oral and Poster Abstracts
Oral
114. Sickle Cell Disease, Sickle Cell Trait, and Other Hemoglobinopathies, Excluding Thalassemias: Clinical and Epidemiological: Tools and Biomarkers for Improving Sickle Cell Disease Treatments

Research, Sickle Cell Disease, Adult, Translational Research, Assays, Clinical Research, Hemoglobinopathies, Pediatric, Diseases, Real-world evidence, Technology and Procedures, Study Population, Human

Amira Idrizovic, MSc, PhDc^1,2^*, Marissa J.M. Traets, MD³^*, Anna Collado Gimbert, MD^1,4^*, Sara Reidel, MSc¹^*, Sigrid van der Veen, MSc⁵^*, Anne-Laure Pham Hung D'Alexandry D'Orengiani, PhD⁶^*, Mirco D'Agnolo, MS⁷^*, Emmanuel Adu, MSc⁸^*, Mickael MARIN, Engineering degree⁹^*, Emmanuelle Bourdelier, Technician¹⁰^*, Elisabetta Mezzalira, RN, MS, PhDc¹¹^*, Betzabel Cajiao Garcia, MSc¹²^*, Santiago Zazo, Prof., PhD¹³^*, Tiziana Sanavia, PhD¹⁴^*, Cesare Rollo, PhD¹⁴^*, David Beneitez, MD^15,16^*, Bart J. Biemond, MD, PhD¹⁷, Erfan Nur, MD, PhD¹⁸, Karin Fijnvandraat, MD, PhD¹⁹^*, Saskia EM Schols, MD, PhD²⁰^*, Anita W. Rijneveld, MD, PhD²¹, Marjon H. Cnossen, MD, PhD²²^*, Aida Kidane, MD^23,24^*, Nicolas Hebert, PhD²⁵^*, Maria Paola Boaro, MD²⁶^*, Slimane Allali²⁷^*, Caroline Le van kim, Prof.²⁸^*, Anna Ruiz Llobet, MD²⁹^*, Béatrice Gulbis, MD, PhD³⁰^*, Federico Alvarez, Prof, PhD³¹^*, Piero Fariselli, PhD¹⁴^*, Petros Kountouris, PhD³²^*, Raffaella Colombatti, MD, PhD³³^*, Pablo Bartolucci, MD, PhD³⁴^*, Mariane De Montalembert, MD, PhD³⁵, Richard van Wijk, PhD³⁶, Eduard J. Van Beers, MD, PhD³⁷, Maria del Mar Mañú-Pereira, PhD¹^* and Minke A.E. Rab, MD, PhD^3,38

¹Rare Anemia Disorders Research Laboratory, Cancer and Blood Disorders in Children, Vall d'Hebron Research Institute / Vall d'Hebron University Hospital, Barcelona, Spain
²Genetics and Microbiology department, Universitat Autonoma de Barcelona, Barcelona, Spain
³Central Diagnostic Laboratory - Research, University Medical Center Utrecht, Utrecht, Netherlands
⁴Pediatric Oncology and Hematology, Vall d'Hebron Barcelona Hospital, Barcelona, Spain
⁵Center for Benign Hematology, Thrombosis and Hemostasis - Van Creveldkliniek, University Medical Center Utrecht, Utrecht, Netherlands
⁶University Paris-Est-Créteil, IMRB, Inserm U955, Laboratory of excellence LABEX, Henri Mondor University Hospitals, APHP, Sickle Cell Referral Center-UMGGR, Créteil, FRA
⁷Department of Women's and Child's Health, University of Padova, Padova, Italy
⁸University Paris-Est-Créteil, IMRB, Inserm U955, Laboratory of excellence LABEX, Henri Mondor University Hospitals, APHP, Sickle Cell Referral Center-UMGGR, Creteil, France
⁹Necker, Université Paris Cité, INSERM, BIGR, PARIS, FRA
¹⁰Institut de Recherche Saint Louis, Université Paris Cité, Inserm, CNRS, Paris, France
¹¹Pediatric Hematology, Oncology and Stem Cell Transplant Division, Maternal and Child Health Department, Università degli Studi di Padova, Padova, Italy
¹²Center for Benign Hematology, Thrombosis and Hemostasis - Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, University Medical Center Utrecht, Utrecht, Netherlands
¹³Centro de I+D+i en Procesado de la Información y Telecomunicaciones (IPTC), Universidad Politécnica de Madrid, Madrid, Spain
¹⁴Computational Biomedicine Unit, Department of Medical Sciences, University of Torino, Turin, Italy
¹⁵Vall d’Hebron Hospital Universitari and General Hospital, Barcelona, ESP
¹⁶Hematology Department, Vall d'Hebron Hospital Universitari/Vall d'Hebron Research Institute (VHIR)/Vall d'Hebron Institute of Oncology (VHIO)/Universitat Autònoma de Barcelona (UAB)/ERN-EuroBloodNet, Barcelona, Spain
¹⁷Department of Hematology, Amsterdam University Medical Centers, Amsterdam, Netherlands
¹⁸Department of Hematology, Amsterdam UMC location University of Amsterdam, Amsterdam, Netherlands
¹⁹Department of Pediatric Hematology, Amsterdam UMC, Emma Children's Hospital, Amsterdam, Netherlands
²⁰Department of Hematology, Radboud University Medical Center, Nijmegen, Netherlands
²¹Department of Hematology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, Rotterdam, Netherlands
²²Department of Pediatric Hematology and Oncology, Erasmus MC Sophia Children’s Hospital, Erasmus University Medical Center, Rotterdam, Netherlands
²³Department of Pediatric Hematology, Erasmus MC Sophia Children's Hospital, Rotterdam, Netherlands
²⁴Section on Social and Cognitive Developmental Neuroscience, National Institutes of Mental Health, Bethesda, MD
²⁵University Paris-Est-Créteil, IMRB, Inserm U955, Laboratory of excellence LABEX, Établissement Français du Sang (EFS), Créteil, France
²⁶Department of Women’s and Children’s Health, Padova University, Padova, Italy
²⁷Sickle Cell Center, Pediatrics Department, Necker Hospital for Sick Children, APHP, Paris Cité University, Paris, FRA
²⁸Necker Hospital, University Paris Cité/Inserm, Paris, France
²⁹Pediatric Hematology, Hospital Sant Joan de Déu, Barcelona, Barcelona, ESP
³⁰Centre of Human genetics, Hôpital Universitaire de Bruxelles, Université Libre de Bruxelles, Brussels, Belgium
³¹Universidad Politécnica de Madrid, Madrid, Spain
³²Molecular Genetics Thalassaemia Department, The Cyprus Institute of Neurology and Genetics, Nicosia, CYP
³³Department for Woman's and Child's Health, University of Padova, Padova, Italy
³⁴University Paris-Est-Créteil, IMRB, Inserm U955,, Henri Mondor University Hospitals, APHP, Sickle Cell Referral Center-UMGGR, Creteil, France
³⁵Department of General Pediatrics and Pediatric Infectious Diseases, Sickle Cell Center, Necker-Enfants Malades Hospital, Neuilly, France
³⁶Central Diagnostic Laboratory, University Medical Center Utrecht, Utrecht, Netherlands
³⁷Center for Benign Hematology, Thrombosis and Hemostasis - Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, Utrecht, Netherlands
³⁸Department of Hematology, Erasmus MC, Rotterdam, Netherlands

Introduction: Sickle cell disease (SCD) is a hereditary disorder characterized by the production of structurally abnormal hemoglobin S (HbS) in red blood cells (RBCs). Under low oxygen saturation, HbS polymerizes, causing RBCs to deform, leading to hemolytic anemia, recurrent vaso-occlusive episodes (VOE) and organ damage. VOE are unpredictable and result in long-term morbidity and early mortality.

RBC deformability and sickling tendency can be assessed ex vivo using oxygen gradient ektacytometry (oxygenscan). Key patient specific parameters are RBC deformability at normoxia (EImax), deformability upon deoxygenation (EImin), and pO2 at which sickling is initiated (PoS). In this study we developed two novel parameters: Slope that reflects how rapidly RBCs sickle during deoxygenation and the EI20 that is RBC deformability measured at a fixed pO2 of 20mmHg. Within GenoMed4ALL project clinical and laboratory data is integrated with oxygenscan parameters to enable early recognition of disease severity and individualize treatment options, addressing a critical need for precision medicine in SCD.

Aim: The aim is to explore how 2 novel parameters (Slope, EI20) perform compared to key oxygenscan parameters (EImin, EImax, PoS) in a multi-national cohort study in regard to correlations with laboratory markers of SCD severity.

Methods: Oxygenscan data was obtained for 807 SCD (HbSS, HbSβ0, HbSC, HbSβ+) patients at different EU health centres or consortia: 5 centers of SCORE consortium, The Netherlands (n=193), 8 Spanish health centers (n=169), Hospital Necker Paris (n=117), Hospital Henry Mondor (n=217) and University of Padova (n=111). All patients were older than 2 years and in steady state (no transfusions nor acute events <3 months) at the time of oxygenscan analysis. Oxygenscan measurements were performed in duplicates using the Laser-Optical Rotational Red Cell Analyzer (RR Mechatronics). Pearson’s test was used to assess linear correlations between laboratory parameters and oxygenscan values and groups were compared using a t-test or ANOVA.

Results: In this preliminary analysis, we evaluated oxygenscans from 350 patients with complete laboratory and clinical data. PoS was significantly lower in HbSβ0 (23,9, n=30) compared to HbSS (33,2, n=240, p<0.001). In contrast we found a lower EImax (0.41) compared to HbSS (0.46, p<0.05). The 2 novel parameters Slope and EI20 were significantly lower in patients with HbSS genotype (p<0.001) compared to other patients with HbSβ0, HbSC and HbSβ+ genotypes.

Inflammatory markers (neutrophils and CRP) showed significant correlations solely in HbSβ0 patients with oxygenscan parameters. CRP positively correlated with the PoS (r=0.416, p<0.05) and neutrophils positively correlated with EImin, PoS, and EI20. Hemolysis markers such as bilirubin correlated with EImax (r=-0.403, p<0.05), LDH correlated with EImax (r=0.602, p<0.01), EI20 (r=0.512, p<0.01 and Slope (r=-0.389, p<0.05) in HbSβ0. In contrast, bilirubin and LDH correlated solely with Slope (r=0.201 and r=-0.219, p<0.01) and unconjugated bilirubin with EI20 (r=0.152, p<0.05) and Slope (r=0.201, p<0.001) in HbSS. All oxygenscan parameters were significantly correlated with hemoglobin (all p<0.001), reticulocyte count (all p<0.01), and fetal hemoglobin (all p<0.05) in HbSS patients which is in accordance with previous findings.

Conclusion: Oxygenscan is a rapid and reproducible technique that quantifies sickling behaviour of RBCs. HbSβ0 patients are often grouped with HbSS patients due to the comparable clinical phenotype, however, our findings regarding RBC function suggest they might present specific different characteristics. We here introduce two novel parameters: slope and EI20. Slope that reflects heterogeneity of the RBC population and EI20 were found to be correlated to both hemolysis and inflammatory markers in HbSS and HbSβ0 highlighting the potential of these 2 parameters. Ongoing studies aim to explore how laboratory data and clinical complications of all patients within the GenoMed4ALL project are associated with oxygen gradient ektacytometry by implementing a generalized hierarchical linear mixed model. This approach will improve our understanding of SCD pathophysiology and enable personalized medicine in SCD.

Disclosures: Idrizovic: Agios: Research Funding; Bristol Myers Squibb: Research Funding; Novartis: Research Funding. Traets: Agios Pharmaceuticals: Research Funding. Collado Gimbert: Novartis: Research Funding; Agios: Research Funding; Bristol myers squibb: Research Funding; Novonordisk: Consultancy; Pfizer: Consultancy. Reidel: Agios: Research Funding; Bristol Myers Squibb: Research Funding; Novartis: Research Funding. Beneitez: Vertex Pharmaceuticals: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novo Nordisk: Consultancy, Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb (Celgene): Consultancy, Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Sobi: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Agios: Consultancy, Membership on an entity's Board of Directors or advisory committees. Biemond: pfizer: Consultancy, Research Funding; sanofi: Honoraria; novartis: Research Funding; BMS: Consultancy, Research Funding; novo nordisk: Honoraria. Nur: Novartis: Research Funding; vertex: Speakers Bureau. Fijnvandraat: NovoNordisk: Consultancy, Research Funding; Roche: Consultancy; CSL Behring: Research Funding; ISTH SSC: Membership on an entity's Board of Directors or advisory committees; Sanofi: Consultancy; SOBI: Consultancy, Research Funding. Rijneveld: Vertex: Other: Advisory board. Cnossen: Takeda: Research Funding, Speakers Bureau; Pfizer: Research Funding, Speakers Bureau; Bayer: Research Funding, Speakers Bureau; CSL Behring: Research Funding, Speakers Bureau; Novo Nordisk: Research Funding, Speakers Bureau; Novartis: Research Funding, Speakers Bureau; Nordic Pharma: Research Funding, Speakers Bureau; Roche: Membership on an entity's Board of Directors or advisory committees; Bayer: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees. Kidane: CSL Behring: Research Funding. Gulbis: Novartis: Research Funding; Agios Pharmaceutical: Research Funding; Bristol-Myers Squibb SA: Research Funding. Kountouris: Agios: Research Funding. Colombatti: Forma Therapeutics: Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Vertex/Addmedica: Membership on an entity's Board of Directors or advisory committees; Agios: Membership on an entity's Board of Directors or advisory committees; Novo Nordisk: Membership on an entity's Board of Directors or advisory committees. Bartolucci: Addmedica: Consultancy, Other: member advisory board; Pfizer: Consultancy; Emmaus: Consultancy; Innovhem: Other: Founder; Novartis: Consultancy, Other: member advisory board and member steering commitee; Bluebird: Consultancy; Roche: Consultancy; JazzPharma: Consultancy. De Montalembert: Theravia: Consultancy, Membership on an entity's Board of Directors or advisory committees; novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; vertex: Membership on an entity's Board of Directors or advisory committees. van Wijk: Pfizer: Research Funding; RR Mechatronics: Consultancy; Agios Pharmaceuticals: Research Funding. Van Beers: Agios Pharmaceuticals, Inc.: Consultancy, Research Funding. Mañú-Pereira: Bristol Myers Squibb: Research Funding; Agios: Other: Advisory board, Research Funding; Novartis: Research Funding. Rab: Agios: Research Funding; RR Mechatronics: Research Funding; Pfizer: Research Funding.

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290 Oxygen Gradient Ektacytometry Is Associated with Markers of Hemolysis and Inflammation in a Large Sickle Cell Disease Cohort within the GenoMed4ALL Project